J Periodontal Implant Sci.  2019 Aug;49(4):215-227. 10.5051/jpis.2019.49.4.215.

Periodontal healing using a collagen matrix with periodontal ligament progenitor cells in a dehiscence defect model in beagle dogs

• Yoo SY
• Lee JS
• Cha JK
• Kim SK
• Kim CS

• KMID: 2455799
• DOI: http://doi.org/10.5051/jpis.2019.49.4.215

Abstract

PURPOSE
To histologically characterize periodontal healing at 8 weeks in surgically created dehiscence defects in beagle dogs that received a collagen matrix with periodontal ligament (PDL) progenitor cells.
METHODS
The bilateral maxillary premolars and first molars in 6 animals were used. Standardized experimental dehiscence defects were made on the buccal side of 3 premolars, and primary culturing of PDL progenitor cells was performed on the molars. Collagen matrix was used as a scaffold and a delivery system for PDL progenitor cells. The experimental sites were grafted with collagen matrix (COL), PDL progenitor cells with collagen matrix (COL/CELL), or left without any material (CTL). Histologic and histomorphometric analyses were performed after 8 weeks.
RESULTS
The defect height from the cementoenamel junction to the most apical point of cementum removal did not significantly differ across the CTL, COL, and COL/CELL groups, at 4.57±0.28, 4.56±0.41, and 4.64±0.27 mm (mean ± standard deviation), respectively; the corresponding values for epithelial adhesion were 1.41±0.51, 0.85±0.29, and 0.30±0.41 mm (P<0.05), the heights of new bone regeneration were 1.32±0.44, 1.65±0.52, and 1.93±0.61 mm (P<0.05), and the cementum regeneration values were 1.15±0.42, 1.81±0.46, and 2.57±0.56 mm (P<0.05). There was significantly more new bone formation in the COL/CELL group than in the CTL group, and new cementum length was also significantly higher in the COL/CELL group. However, there were no significant differences in the width of new cementum among the groups.
CONCLUSIONS
PDL progenitor cells carried by a synthetic collagen matrix may enhance periodontal regeneration, including cementum and new bone formation.

Keyword

Collagen matrix; Dehiscence defect; Periodontal ligament progenitor cells; Periodontal regeneration

MeSH Terms

Animals
Bicuspid
Bone Regeneration
Collagen*
Dental Cementum
Dogs*
Molar
Osteogenesis
Periodontal Ligament*
Regeneration
Stem Cells*
Tooth Cervix
Transplants
Collagen

Figure

• 

  Figure 1 Clinical photographs of the surgical procedure. (A) Preoperative view of the first, second, and third premolars. (B) A dehiscence defect with a size of 5×5 mm (width × length) was surgically created. (C) Each site of the dehiscence defect was grafted with collagen matrix with PDL progenitor cells (COL/CELL group), grafted with collagen matrix alone (COL group), or left without any material (CTL group). (D) Primary closure was then performed. COL group: collagen matrix group, COL/CELL group: PDL progenitor cells with collagen matrix group, PDL: periodontal ligament, CTL group: sham control group.

• 

  Figure 2 Schematic drawing of measurements. The bottom of the defect was set at the most apical point of the planed root surface, the most coronal point of newly formed cementum on the root surface was marked by a straight line, and the most apical point of the long junctional epithelium was also marked by a straight line. The newly formed bone was distinguished from the preexisting alveolar bone by its color. B: bone, PDL: periodontal ligament, C: cementum, JE: junctional epithelium, DH: defect height, NC: new cementum, CTA: connective-tissue adhesion, EA: epithelial adhesion, NB: new bone.

• 

  Figure 3 Characterization of PDL progenitor cells as mesenchymal stem cells. (A, B) Colony-forming-units assay revealed colony formation at 14 days after cell plating (stained with crystal violet). Although the colony formation decreased from (A) P3 to (B) P4, the size of the colonies was similar at (C) P3 and (D) P4. Adipogenic and osteogenic differentiation were induced after 4 and 2 weeks, respectively. PDL: periodontal ligament; P: passage.

• 

  Figure 4 Scanning electron microscopy photographs showing collagen matrix with periodontal ligament progenitor cells in the experimental group at observation periods of (A) 0 minutes, (B) 10 minutes, and (C) 30 minutes . The number of cells tended to decrease from 0 to 30 minutes, with some cells showing a spindle-like appearance.

• 

  Figure 5 Photomicrographs of the buccolingual section in the dehiscence defects of periodontal tissue in the (A) CTL, (B) COL, and (C) COL/CELL groups, displaying the sites from the apical extension of the surgically created defect (apical insert; a, d, and g) along the root surface to the coronal extension of the newly formed alveolar bone and cementum (midroot insert; b, e, and h), and the apical extension of epithelial adhesion (c, f, and i; hematoxylin-eosin stain). PDL: periodontal ligament, AB: alveolar bone, Ce: cementum, JE: junctional epithelium, COL group: collagen matrix group, COL/CELL group: PDL progenitor cells with collagen matrix group, CTL group: sham control group.

• 

  Figure 6 Low-magnification photographs of the buccolingual section of the dehiscence defects of periodontal tissue in the (a) CTL, (b) COL, and (c) COL/CELL groups, along with high-magnification photographs from the sites of the dehiscence defects of periodontal tissue in the (A) CTL, (B) COL, and (C) COL/CELL groups. The newly formed PDL did not differ across the 3 groups (hematoxylin-eosin stain). Ce: cementum, PDL: periodontal ligament, AB, alveolar bone, CTL group: sham control group, COL group: collagen matrix group, COL/CELL group: PDL progenitor cells with collagen matrix group.

• 

  Figure 7 Comparison of periodontal tissue formation. (A) The defect height did not differ significantly across the 3 groups (P=0.84). (B) New bone formation did not differ significantly across the 3 groups (P=0.058). However, the Tukey post hoc test revealed a significant difference between the CTL and COL/CELL groups (P=0.046). (C) New cementum formation differed significantly across the 3 groups (P=0.001). CTL group: sham control group, COL group: collagen matrix group, COL/CELL group: PDL progenitor cells with collagen matrix group. a)P<0.05.

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